Sex preselection by flow cytometric separation of X and Y chromosome-bearing sperm based on DNA difference: a review

La, Johnson

doi:10.1071/rd9950893

Cited by 91 publications

(44 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Isolations were performed using a flow cytometer with specific modifications as developed by Johnson et al (13). Modifications included an additional fluorescence detector positioned at a right angle (90 o ) and a novel sample injection tip designed to "orient" the spermatozoa to the laser beam (14,15). Fluorescent signal from Hoechst-stained spermatozoa oriented with their brightest edge toward the 90 o detector was collected by 0 o detector and showed bimodal distribution.…”

Section: Resultsmentioning

confidence: 99%

CAG Repeat Lengths in X- and Y-bearing Sperm Indicate That Gender Bias during Transmission of Huntington's Disease Gene Is Determined in the Embryo

Kovtun

Welch

Guthrie

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

The size of the CAG tract at the Huntington's disease (HD) locus upon transmission depends on the gender of the parent. However, the basis for the parent-of-origin effect is unknown. To test whether expansion and contraction in HD are "imprinted" in the germ cells, we isolated the X-and Y-bearing sperm of HD transgenic mice. Here we show that CAG repeat distributions in the X-and Y-bearing spermatozoa of founding fathers do not differ. These data show that gender-dependent changes in CAG repeat length arise in the embryo.The mechanism for expansion in HD 1 and other trinucleotide disorders is not known. However, one of the poorly understood features of expansion is the gender bias that is associated with transmission. Large changes in repeat number are known to be transmitted through the paternal line in HD (1, 2), SCA1 (3), dentatorubral-pallidoluysian atrophy (DRPLA) (4), MachadoJoseph disease (5), and spinal and bulbar muscular atrophy (SBMA) (6) and through the maternal line in Fragile X (7, 8). The gender bias transmission has also been documented in transgenic mice (9 -12). Similar to humans, HD transgenic mice transmit expansions predominantly through the male germ line (9,10,12). Moreover, we found that expansion and contraction of CAG repeat size in the hHD transgene depended on the gender of the progeny (9).Expansions are primarily seen in males and contractions dominate in females. The molecular basis for the parent-oforigin effect is unknown. The gender dependence of expansion can be due to differences in CAG repeat distribution in X-and Y-bearing sperm. Alternatively, the change in repeat number, contraction, or expansion can take place after fertilization in early embryogenesis. In order to distinguish between these two mechanisms, the isolation and purification of the X-and Ybearing sperm were required. This, however, was not possible using conventional technology.In present work, we have developed techniques and used advanced technology to separate pure populations of X-and Y-bearing germ cells from HD transgenic mice. We show here that CAG repeat distributions in X-and Y-bearing parental sperm are not different. Repeat expansion occurs equally well within both X and Y chromosomes. This indicates that the gender-dependent processing of CAG repeats must take place post-zygotically. Thus, these data provide evidence for a new kind of imprinting that may be important in the interpretation of genetic data in other systems. EXPERIMENTAL PROCEDURESAnimals-HD transgenic male mice (line B6CBA-TgN R6/1) were originally purchased from The Jackson Laboratory. The colony was maintained at the animal core facility, Mayo Clinic/Foundation. Animals were routinely screened for the presence of HD transgene by PCR (10, 18).Preparation of Single Cell Suspension and Flow Sorting-Mouse epididymis was dissected from the animal, placed in phosphate-buffered saline, and chopped with a razor. Resulting suspension was filtered through cheesecloth. The supernatant was centrifuged at 1500 rpm, and the pellet was resuspended...

show abstract

Section: Resultsmentioning

confidence: 99%

CAG Repeat Lengths in X- and Y-bearing Sperm Indicate That Gender Bias during Transmission of Huntington's Disease Gene Is Determined in the Embryo

Kovtun

Welch

Guthrie

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…laser impact effect (Johnson 1995, Maxwell et al 1997, Maxwell & Johnson 1999, Parrilla et al 2001. In truth, a reduction in the fertility index without any increase in lost pregnancies, abortions or morphological abnormalities should be obtained as observed in our experiments where, in addition, the cytogenetic analysis did not shown any deviations from baseline levels according to the phenotypical normalcy of the offspring.…”

Section: Discussionmentioning

confidence: 99%

Hoechst 33342 stain and u.v. laser exposure do not induce genotoxic effects in flow-sorted boar spermatozoa

Parrilla¹,

Vázquez²,

Cuello³

et al. 2004

Reproduction

View full text Add to dashboard Cite

Sex selection by flow cytometry/cell sorting involves the staining of spermatozoa with Hoechst 33342 in combination with the impact of a u.v. laser beam, two potentially mutagenic agents. A phenotypic and cytogenetic study of lymphocytes of piglets born after insemination with spermatozoa stained with Hoechst 33342 and from piglets obtained from stain-sorted spermatozoa was performed to evaluate the genotoxic effect of Hoechst 33342 staining and u.v. laser irradiation on the offspring. Lymphocytes from piglets born after insemination with unstained spermatozoa, but from the same ejaculate, were used as a control group. Peripheral blood lymphocytes from these piglets were cultured following a standard cell culture protocol. Cells were then collected by centrifugation, subjected to hypotonic solution and fixed and dropped onto slides. Sister chromatid exchanges (SCEs) and chromosome aberrations (CAs: including chromosome and chromatid breaks) per cell were scored in 50-s division metaphase spreads from each donor. Reproductive parameters and litter performance of all inseminations performed were also recorded in all groups. Data were analyzed by ANOVA. No significant increase (P > 0.05) of SCE and CA frequencies were observed in piglets born from stained spermatozoa or from stain-sorted spermatozoa with respect to controls (untreated sperm). The results indicated that no mutagenic effect on spermatozoa, expressed as increases in the incidence of abnormalities in the resulting offspring and also as increases in SCE and CA frequencies on lymphocytes from these individuals, was induced by the staining of boar spermatozoa with Hoechst 33342, nor by combination of staining with laser impact during flow cytometry.

show abstract

“…Many offspring have successfully been produced from sperm sorted by flow cytometer followed by surgical insemination [3,4], IVF [7,8,58], and ICSI [5,6]. Before development of high-speed sorting technology [35,36], the primary limiting factor for use of sperm sorted by flow cytometer in artificial insemination is that only small numbers of sperm were available in a given time period [59]; this should be overcome because good motility and a large number of sorted sperm are required for calf production.…”

Section: Production Of Sex Preselected Offspring With Flow Sorted X-amentioning

confidence: 99%

Sex Preselection in Bovine by Separation of X- and Y-Chromosome Bearing Spermatozoa

Hamano

2007

Journal of Reproduction and Development

View full text Add to dashboard Cite

Abstract. Flow cytometrically-sorted sperm has been involved in the production of sex preselected offspring. More than 30,000 bovine offspring have been produced using AI and other means using spermatozoa separated by flow cytometer. Flow cytometric sperm sorting based on differences in their DNA content is the best method for separation of X-and Y-chromosome bearing spermatozoa. At first, flow cytometers were modified for DNA confirmation and sorting of sperm with high resolution. The beveled insertion needle can regulate orientation of flat-shaped bull sperm heads. The forward fluorescence detector is essential for measuring the DNA content of sperm. Recently, high-speed sperm sorting with orienting nozzles has resulted in production of 90% pure X-and Ysperm at rate of 15-20 million sperm per hour. Application of this new technique will enable conduct of more conventional technologies for both artificial insemination and cryopreservation in the bovine and in other farm animals using X-or Y-sperm.

show abstract

Sex preselection by flow cytometric separation of X and Y chromosome-bearing sperm based on DNA difference: a review

Cited by 91 publications

References 0 publications

CAG Repeat Lengths in X- and Y-bearing Sperm Indicate That Gender Bias during Transmission of Huntington's Disease Gene Is Determined in the Embryo

CAG Repeat Lengths in X- and Y-bearing Sperm Indicate That Gender Bias during Transmission of Huntington's Disease Gene Is Determined in the Embryo

Hoechst 33342 stain and u.v. laser exposure do not induce genotoxic effects in flow-sorted boar spermatozoa

Sex Preselection in Bovine by Separation of X- and Y-Chromosome Bearing Spermatozoa

Contact Info

Product

Resources

About